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Cloning Technologies

  • Andrew Fontes
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 997)

Abstract

One major obstacle in realizing the potential behind human embryonic stem cells (hESC) is the availability of efficient and reliable engineering methods. Such methods require cloning technologies that can be applied to a variety of platforms and can serve multiple functions. In the last two decades cloning technologies have become more efficient, widening the bottleneck in creating engineered hESC lines. Using TOPO® TA cloning kits, genes can be efficiently amplified and inserted into target vectors with minimal manipulation and purification. For more complex cloning procedures we introduce the Multisite Gateway® system. This is a cloning platform based on integrase technology that allows for the generation of complex multicistronic gene configurations that can transverse a variety of platforms with ease. These technologies allow the end user to quickly and efficiently select clones, as well as combine multiple genetic elements of interest between platform technologies in a high-throughput manner, providing scientists with a toolbox to create tools to dissect stem cell biology.

Key words

Gateway® TOPO® hESC Cloning Engineering 

References

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Andrew Fontes
    • 1
  1. 1.Primary and Stem Cell SystemsLife TechnologiesCarlsbadUSA

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